EP1335701B1 - Composition anti-adipeuse a base d'extraits de bulbes d'ail et utilisations cosmetiques et therapeutiques - Google Patents

Composition anti-adipeuse a base d'extraits de bulbes d'ail et utilisations cosmetiques et therapeutiques Download PDF

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Publication number
EP1335701B1
EP1335701B1 EP01982561A EP01982561A EP1335701B1 EP 1335701 B1 EP1335701 B1 EP 1335701B1 EP 01982561 A EP01982561 A EP 01982561A EP 01982561 A EP01982561 A EP 01982561A EP 1335701 B1 EP1335701 B1 EP 1335701B1
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Prior art keywords
allium sativum
active agent
adipose
composition
composition according
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German (de)
English (en)
French (fr)
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EP1335701A2 (fr
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Isabelle Tomatis
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Pierre Fabre Dermo Cosmetique SA
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Pierre Fabre Dermo Cosmetique SA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • A61Q19/06—Preparations for care of the skin for countering cellulitis
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/92—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof
    • A61K8/922—Oils, fats or waxes; Derivatives thereof, e.g. hydrogenation products thereof of vegetable origin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
    • A61K8/9783—Angiosperms [Magnoliophyta]
    • A61K8/9794—Liliopsida [monocotyledons]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/04—Anorexiants; Antiobesity agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/06—Antihyperlipidemics

Definitions

  • the invention relates to a topical anti-adipose treatment composition, and the uses of this composition and / or extracts of garlic bulbs for cosmetic purposes, for the preventive or curative treatment of cellulite and localized dermal fat overloads, or therapeutically for the treatment of obesity.
  • cellulite is the accentuation of adipose tissue in certain areas of the body, especially on the hips, buttocks, knees and forearms. This adipose tissue or fat mass of the body is present under almost the entire surface of the skin. Cellulite is often associated with an overall fat overload and the stability of the fat mass is an important point in controlling the development of cellulite.
  • adipocyte is a large cell in which 80% of the volume consists of one or more vacuoles of lipids. Its size is variable. If the vacuole is overloaded with fat, the volume of the adipocyte increases and the dermal connective tissue thickens.
  • Cellulite also depends on functional criteria, because it is also a localized cluster, associated with a deficiency of the venous tone and / or an alteration of the lymphatic system.
  • the first deformation of the cellulite tissue takes place at the level of the adipocytes. As soon as the volume of the adipocytes increases, the septa will undergo a deep stretch which appears on the surface of the epidermis under the form of depressions in some places. These depressions appear even more if the skin is pinched: it is the orange peel.
  • the second deformation is at the level of the fundamental substance.
  • the fundamental substance is modified according to the quantity of water that it retains. This explains the loss of flexibility of the skin on a cellulite location. The water pressure retained by the basic substance will crush the cells and thereby cause the degradation of elastin and collagen fibers.
  • the adipocyte functions as a reservoir of energy for the body.
  • the adipocyte is able to store energy in the form of triglycerides in its lipid droplets (this is lipogenesis) and then release this energy in the form of fatty acids in the circulation. blood (it is lipolysis).
  • the lipogenic function of the adipocyte requires the presence of fatty acids essentially derived from circulating lipoproteins rich in triglycerides.
  • lipolysis corresponds to the separation of the constituents of adipocyte triglycerides into glycerol on the one hand, and fatty acids on the other hand.
  • This lipolysis is done under the action of a limiting enzyme, the hormone-sensitive lipase.
  • Activation of the hormone-sensitive lipase depends on the cyclic AMP content.
  • hormones and various agents are capable of modulating intracellular cAMP concentrations, and therefore participate in the control of hormone-sensitive lipase therefore to lipolysis.
  • all known slimming creams contain at least one active agent that promotes lipolysis and / or inhibits lipogenesis, or that aims to restore capillary and lymphatic microcirculation.
  • hypertrophy The excessive development of the adipose mass is usually related to an increase in the volume of adipocytes, and their content in triglycerides without there being an increase in their number: it is called hypertrophy. In some cases of excess weight, hypertrophy is accompanied by hyperplasia, that is to say, an increase in the number of fat cells.
  • the preadipocyte is a thin cell, fibroblastic morphology: it is he and not the adipocyte, which is able to multiply.
  • the terminal adipocyte differentiation is done in two stages with transformation of the preadipocyte into mature adipocyte, the latter being able to accumulate triglycerides.
  • the present invention aims to fill this gap and to main objective of providing a therapeutic or cosmetic composition for the treatment by topical application anti-adipose externally (on the skin), in particular acting very effectively on blocking the differentiation of preadipocytes into adipocytes.
  • the invention aims to provide an improved cosmetic or therapeutic treatment of cellulite, localized dermal fat overloads, and obesity.
  • the invention further aims to propose new uses for cosmetic or therapeutic purposes, extracts-especially non-aqueous extracts-of Allium sativum (garlic) bulbs.
  • the term “bulbs” includes both plant bulbs (clusters of garlic) and garlic cloves separated from each other, peeled or not.
  • the invention also aims at proposing new compositions based on garlic bulb extracts and their applications, in particular for cosmetic or therapeutic purposes.
  • Another object of the invention is to provide a therapeutic composition acting on mature hypertrophied adipocytes in the context of the treatment of obesity.
  • extracts extract, essential oil, absolute, concrete, oleoresin .
  • composition according to the invention comprises an amount of at least one anti-adipose active agent adapted to at least substantially inhibit the differentiation of preadipocytes into mature adipocytes, without substantially causing irritation or skin sensitization.
  • the composition comprises an effective amount of at least one anti-adipose active agent chosen from non-aqueous extracts of Allium sativum bulbs.
  • the amount of extract (s) of Allium sativum bulbs is between 3 ppm and 20 ppm - in particular of the order of 10 ppm-.
  • the composition comprises, as active anti-adipose agent, an essential oil of Allium sativum bulbs and an emulsifying agent.
  • the essential oil / emulsifying agent mixture is incorporated in a gel and a liquid excipient such as water.
  • the composition comprises, as active anti-adipose agent, at least one absolute of Allium sativum bulbs in aqueous solution.
  • it comprises at least one absolute obtained from a concrete extracted from Allium sativum bulbs.
  • it also comprises at least one absolute obtained from an extraction oleoresin extracted from Allium sativum bulbs.
  • composition according to the invention is advantageously characterized in that it comprises, as an anti-adipose active agent, at least one extract of Allium sativum bulbs chosen from an essential oil, an absolute obtained from of a concrete extracted with hexane, an absolute obtained from a concrete extract extracted with ethyl acetate, an absolute obtained from an extraction oleoresin extracted with acetone.
  • the amount thereof is advantageously between 3 ppm and 20 ppm -particularly of the order of 10 ppm-.
  • the amount of extract (s) is advantageously between 5 ppm and 20 ppm - in particular of the order of 10 ppm-.
  • the proportion of absolute (s) in ppm of the various compositions according to the invention refers as to at a quantity of absolute (s) "pure (s)", that is to say an absolute containing less than 2% residual ethanol.
  • the invention is thus based on the very surprising and unexpected discovery that extracts of Allium sativum bulbs exhibit a very effective activity of blocking the differentiation of fat cells.
  • Allium sativum is known for many centuries, as having antiseptic, anti-viral, cardio-protective, tonic, diuretic.
  • EP-0923 937 describes a composition for the treatment of skin disorders such as allergies, depigmentations, mycoses, cracks, hyperkeratoses, cuts, burns, comprising at least two plant extracts in the form of oil or powder obtained by extraction with solvents from dried plants. It is therefore extraction oleoresins (or resinoids).
  • the composition comprises 1 to 3% of an oleoresin extraction of garlic extracted with hexane, which has no anti-adipose activity.
  • GB-1106551 discloses the use of plant extracts of the family Nympheaceae as active agents for the therapeutic treatment of neoplasms and cellulitis.
  • the described compositions also include one or more complementary essential oil (s) of plants of other species, in particular of the family Cruciferaceae (for example mustard oil or horseradish oil) or of the Liliaceae family (eg garlic oil), producing a stabilizing effect of Nympheaceae extracts, and an effect synergistic with these extracts.
  • complementary essential oil for the treatment of cellulite topically, essential oils are dissolved in olive oil. This document indicates that said complementary essential oils have no effect of their own.
  • EP-O333548 discloses stable aqueous emulsions of orange essential oil comprising 1 to 45 parts by weight of essential oil; 0.01 to 1 part by weight of a nonionic emulsifier selected from sucroglycerides and sucroesters; 0.2 to 1.5 parts by weight of xanthan gum, and the balance by weight of water.
  • These emulsions may be used for the preparation of aromatization concentrates of food products, in particular soft drinks and confectionery or concentrates for cosmetics.
  • This document indicates theoretically that all the "essences” of plants (in fact any extracts) can be used in the emulsion, and therefore quotes among them in particular "the essence of garlic". Nevertheless, it gives embodiments only in the case of orange essence. Now, with the proportions indicated, a stable emulsion of essential oil can not be obtained with all the known essential oils, and in particular with the essential oil of garlic. This document therefore does not provide realistic teaching on a composition based on essential oil of garlic.
  • JP-04 338 336 discloses oral obesity treatment tablets comprising garlic extracts obtained from dried garlic and extraction with water or ethanol. This document aims to enhance the well-known hypolipidemic effect of certain garlic extracts in the circulating blood.
  • FR-2804319 published August 3, 2001 discloses a slimming cosmetic composition
  • a plant extract containing ANP necessarily obtained by extraction with water and from the stems, leaves and petals of plants.
  • This document lists various usable plants, including garlic, but does not describe a composition comprising an extract of Allium sativum bulbs.
  • the mechanism of action of the composition is to promote the lipolysis of adipocytes, but in no way acts on the transformation of preadipocytes into mature adipocytes.
  • the extracts of Allium sativum according to the invention act locally on the adipose cell without causing the release into the bloodstream of a portion of its fatty acids which, if not burned, could prove detrimental in terms of health ( deposit on the arterial wall or intra-abdominal tissue).
  • the extracts of Allium sativum according to the invention act on a new side of the fight against cellulite: by limiting the recruitment of dormant preadipocytes they limit the expansion of adipose tissue.
  • an anti-adipose agent according to the invention also ensures good vascularization of the adipose tissue by reducing the levels of vasopressor agents.
  • the extracts of Allium sativum according to the invention thus constitute anti-adipose active agents having maximum effectiveness making it unnecessary to multiply the combinations of active ingredients in cosmetic or therapeutic slimming formulations.
  • anti-adipose active agents according to the invention are also active at mature hypertrophied adipocytes. They thus act on the two processes of fat growth, hyperplasia and hypertrophy, which makes it possible to use them successfully in the therapeutic field for the treatment of obesity.
  • anti-adipose active agents lies in the fact that the phenomena obtained are reversible and therefore they do not lead to a blockage of the physiological mechanism.
  • the therapeutic or cosmetic compositions according to the invention constitute a body care, which preferably is applied externally to areas of the skin to be treated at least once a day, every day of the year.
  • the composition according to the invention requires no other active agent than that according to the invention, and the extract (s) of Allium sativum may be the only agent (s) anti-adipose active agent (s) of said composition. Therefore, the composition according to the invention is free of additional active agent.
  • the (the) extract (s) of Allium sativum bulbs is (are) the sole (s) plant extract (s) of a composition according to the invention.
  • a composition according to the invention can also be used for so-called “flash” treatments consisting of performing a treatment for 2 or 3 months, especially before the summer period.
  • it advantageously comprises a complementary active agent consisting of a lipolytic agent of any type known per se.
  • a composition according to the invention comprises at least one lipolytic agent chosen from lipolytic agents of natural origin (such as caffeine and lipolytic agents) of synthesis (compound (s) of xanthine bases), capable of stimulating the synthesis of cAMP.
  • a composition according to the invention may contain a number of agents capable of: stabilizing the elastic network (for example silicon derivatives); and / or provide an immediate tightening effect (incorporation of synthetic polymers or vegetable proteins (soybean, wheat germ) and / or ensure optimal hydration (incorporation of humectants and / or restructuring agents).
  • agents capable of: stabilizing the elastic network for example silicon derivatives
  • an immediate tightening effect incorporation of synthetic polymers or vegetable proteins (soybean, wheat germ) and / or ensure optimal hydration (incorporation of humectants and / or restructuring agents).
  • composition according to the invention does not require the incorporation of agents capable of improving its penetration power (such as exfoliants).
  • composition according to the invention agents capable of improving cutaneous microcirculation.
  • composition according to the invention may comprise any excipient compatible with the anti-adipose active agents according to the invention, acceptable pharmaceutically or cosmetically, and which may comprise any suitable adjuvants including gelling agents, perfumes, preservatives, stabilizers, colorants, etc. obtain the desired and appropriate dosage form.
  • a composition according to the invention may be free of xanthan gum.
  • an emulsifying agent in particular the product designated Liposol marketed by SIEGEN (Geneva, Switzerland), or any other emulsifier, especially any emulsifier which is neither a sucroglyceride nor a sucroester.
  • the amount of emulsifier used is adapted to obtain the desired emulsion. It is in practice much more important -typically 50 times - than that of essential oil.
  • composition according to the invention is advantageously in the form of a simple lotion.
  • Embodiments of a composition according to the invention are as follows.
  • the essential oil of Allium sativum is first incorporated into an emulsifying agent such as Liposol (SIEGEN, Switzerland), the respective proportions by volume being, for example, order of 0.001% of essential oil, 0.05% of Liposol, and the complement to 100% excipient.
  • an emulsifying agent such as Liposol (SIEGEN, Switzerland)
  • the respective proportions by volume being, for example, order of 0.001% of essential oil, 0.05% of Liposol, and the complement to 100% excipient.
  • a composition according to the invention is extremely simple and inexpensive.
  • the invention extends to a method of cosmetic treatment and / or therapeutic preventive or curative cellulite, localized dermal fat or obesity, characterized in that it is applied topically to areas of skin to treating at least one extract of Allium sativum bulbs as an anti-adipose active agent.
  • Treatment of dermal adipose overloads means treatment of the localized fat mass both in the dermis and in the hypodermis.
  • the invention also relates to a treatment method characterized in that a composition according to the invention is used.
  • the invention also relates to the use of at least one extract of Allium sativum bulbs as an anti-adipose active agent for the preventive or curative cosmetic treatment, by topical application to areas of skin to be treated, the cellulite and localized dermal fat overloads.
  • the invention also relates to the use of at least one extract of Allium sativum bulbs as antiadipose active agent for the preparation of a therapeutic composition for preventive or curative treatment, by topical application to areas of skin to be treated (belly, thighs, hips, buttocks, bulges, limbs ...) of obesity.
  • the invention also relates to a process for the preparation of a therapeutic composition for the preventive or curative treatment, by topical application to areas of skin to be treated, of obesity, characterized in that an effective amount is incorporated into the composition.
  • at least one extract of Allium sativum bulbs as an anti-adipose active agent.
  • an amount of at least one extract of Allium sativum bulbs adapted to cause at least substantially an inhibition of the differentiation of preadipocytes into mature adipocytes and / or for less substantially limit or inhibit the hypertrophy of mature adipocytes, without causing irritation or skin sensitization.
  • the invention also extends to a method of extracorporeal inhibition of the adipocyte conversion of preadipocytes into mature adipocytes, characterized in that an effective amount of at least one extract of Allium sativum bulbs is placed in contact with each other. pre-adipocytes or a biological tissue incorporating them.
  • non-aqueous extract (s) of Allium sativum bulbs are used as active anti-adipose agent (s).
  • At least one essential oil and at least one emulsifying agent are used as the active anti-adipose agent.
  • at least one absolute in aqueous solution is used as the active anti-adipose agent.
  • at least one extract of Allium sativum bulbs chosen from an essential oil is used as active non-adipose agent; an absolute obtained from a concrete extracted with hexane; an absolute obtained from a concrete extracted with ethyl acetate; an absolute obtained from an extraction oleoresin extracted with acetone.
  • a composition according to the invention is used.
  • the invention also relates more generally to a composition adapted for implementing a treatment method (cosmetic and / or therapeutic) or an inhibition method according to the invention.
  • the invention also relates to a composition, a treatment method -particularly cosmetic-a method for preparing a therapeutic composition, a use, characterized in combination by all or part of the characteristics mentioned above or below.
  • Examples 1, 2, 3 and 4 illustrate the biological efficacy on the processes of adipocyte hyperplasia and hypertrophy of an essential oil of Allium sativum bulbs (called HE); an absolute of Allium sativum bulbs obtained from a concrete extracted with hexane (called ACH); an absolute of Allium sativum bulbs obtained from a concrete extract extracted with ethyl acetate (called ACAE); an absolute of Allium sativum bulbs obtained from an extraction oleoresin extracted with acetone (so-called AOA).
  • HE adipocyte hyperplasia and hypertrophy of an essential oil of Allium sativum bulbs
  • ACH an absolute of Allium sativum bulbs obtained from a concrete extracted with hexane
  • ACAE an absolute of Allium sativum bulbs obtained from a concrete extract extracted with ethyl acetate
  • AOA an absolute of Allium sativum bulbs obtained from an extraction oleoresin extracted with acetone
  • a solution of HE introduced into the culture medium is prepared by diluting to 1/10 in water for cell culture (Eurobio®, France), a stock solution of HE (marketed by SIGMA-ALDRICH FINE). CHEMICALS; "Garlic oil, Chinese", ref. W250309) consisting of 18.4 ⁇ l of HE and 981.6 ⁇ l of Liposol (natural emulsifier based on substances extracted from plant cell membranes, marketed by SIEGEN, Switzerland).
  • a control solution introduced into the culture medium is prepared by diluting to 1/10 in water for cell culture (Eurobio®, France), a control mother solution consisting of 18.4 ⁇ l of water for cell culture (Eurobio ®, France) and 981.6 ⁇ l of Liposol.
  • the absolutes according to the invention are prepared according to a process which in itself is conventional, well known and well defined in the perfume industry.
  • an absolute is an extract resulting from at least two extraction steps.
  • the first step is carried out with a volatile non-aqueous extraction solvent (especially non-benzene) and provides a concrete when the starting plant material is fresh or thawed (after being frozen for preservation), or an oleoresin when said material is used in the dry state.
  • the second step is carried out with ethanol as a solvent.
  • the absolute ACH, ACAE and AOA are obtained by a process applied on Allium sativum bulbs.
  • CASA is prepared according to a protocol comparable to that of obtaining the ACH, the first step this time consisting of an extraction with ethyl acetate.
  • ACAE absolute diluted 25% in ethanol, is a clear light brown solution with a medium odor.
  • the AOA is prepared according to a protocol comparable to that of obtaining ACH, the first extraction being this time with acetone, from garlic cloves previously subjected to a drying step. At the end of this first extraction, an extraction oleoresin is obtained. At room temperature, it is in the form of a very dark brown paste.
  • the absolute AOA is in the form of a clear solution of brown color, low odor.
  • the ACH, ACAE and AOA solutions introduced into the culture medium are prepared by diluting 1/10 in water for cell culture (Eurobio®, France) with stock solutions of each absolute (ACH, ACAE or AOA). ) consisting of 18.4 ⁇ l of absolute (ACH, ACAE or AOA) and 981.6 ⁇ l of water for cell culture (Eurobio®, France).
  • the cells are seeded in 6-well plates (seeding density: 15000 cells / well) and maintained until confluence in DMEM-10% SVD medium and antibiotics. At confluence, the preadipocytes are then cultured in DMEM-10% FCS and insulin medium. The treated cells are maintained in the culture medium which is added to the Final 25 ⁇ l of one of the test solutions (HE, or ACH, or ACAE, or AOA) per milliliter of culture medium. The control 3T3-F442A preadipocytes are cultured in the culture medium to which 25 .mu.l of control solution per milliliter of culture medium is added. The culture medium is renewed every 48 hours.
  • test solutions HE, or ACH, or ACAE, or AOA
  • Human preadipocytes (ZenBio®, USA) are grown according to the supplier's instructions. Upon reception, the human preadipocytes are cultured in the appropriate medium provided, to which is finally added 3 .mu.l to 10 .mu.l of one of the test solutions (HE, or ACH, or ACAE, or AOA) per milliliter of medium. culture. The control preadipocytes are kept in culture in the medium alone. The culture medium is renewed every 3 days.
  • solutions of HE, ACH, ACAE or AOA inhibit the process of terminal differentiation of human preadipocytes.
  • the human preadipocytes cultured in the presence of a solution of HE (FIG. 1D), or of a solution of ACH (FIG. 1E) retain a pre-adipocyte morphology: the human cells remain fusiform with very few detectable intra-cytoplasmic lipid droplets .
  • the same results are obtained with ACAE and AOA solutions.
  • the control human preadipocytes (FIG. 1F) lose their fibroblastic morphology, with appearance of a spherical shape and the existence of numerous intra-cytoplasmic lipid vesicles-identifiable by their high refractive intensity.
  • the 3T3-F442A preadipocytes are cultured for 4 days in differentiating medium (DMEM-10% FCS and insulin). Under these conditions, the cells pass to the stage of "mature adipocytes" whose volume and content of triglycerides has been increased. The cells are then treated as before.
  • DMEM-10% FCS and insulin differentiating medium
  • the mature adipocytes treated with a solution of HE (FIG. 2A) have a less rounded cell morphology and a content in lipid vesicles intra-adipocyte strongly decreased, so that the solution of HE, according to the invention is effective in the process of limiting adipocyte hypertrophy, by reversing the differentiation process.
  • the same results are obtained with the solutions of ACH, or ACAE or AOA according to the invention.
  • adipocyte differentiation is an important component of adipose tissue development and obesity.
  • the process of adipocyte differentiation is characterized in vitro by the programmed induction of different genes regulating lipoprotein lipolysis, cell uptake of fatty acids, and synthesis of fatty acids and triglycerides.
  • these genes can be classified as differentiation markers of the type: very early (such as the expression of lipoprotein lipase mRNA), early (such as the expression of PPAR ⁇ 2 mRNA) or late (such as as the expression of leptin) (General Review: Morrison RF, Farmer SR, J. Cell Biochem Suppl.32 / 33: 59-67, 1999).
  • the stage of differentiation of adipocytes can be established precisely by studying the level of expression of the relevant mRNAs (PPAR ⁇ 2 mRNA) or by the level of expression of leptin.
  • PPAR ⁇ nuclear receptors of PPAR type
  • PPAR ⁇ nuclear receptors of PPAR type
  • PPAR ⁇ is strongly expressed in the liver, it regulates the expression of genes involved in lipid metabolism.
  • the expression of PPAR ⁇ is ubiquitous; the function of this receiver remains to be determined.
  • PPAR ⁇ exists as two PPAR ⁇ and PPAR ⁇ 2 isoforms. It is PPAR ⁇ 2 which is strongly expressed in the adipose tissue of mammals. When PPAR ⁇ 2 is activated, it induces the transcription of several adipocyte genes encoding proteins and enzymes involved in the creation and maintenance of the adipocyte phenotype. It plays a key role in the differentiation and metabolism of fat cells.
  • the effect on the expression of PPAR ⁇ 2 mRNA of the solutions of: HE, ACH, ACAE or AOA was determined after 3 days (FIG. 3A, 3T3-F442A cells) or 6 days (FIG. human adipocytes) of treatment.
  • the content of PPAR ⁇ 2 mRNA is analyzed by RT-PCR (Reverse Transcription Polymerase Chain Reaction), after having previously: purified the RNA according to the RNeasy Total RNA System kit procedure (Qiagen®), quantified the RNA content samples by the measurement and the ratio of absorbances at 260 and 280 nm, and verified after migration on agarose gel the quality of the RNAs obtained.
  • the complementary DNA is synthesized from 1 ⁇ g of total RNA (3T3-F442A) or 0.2 ⁇ g of total RNA (human pre-adipocytes).
  • the amplifications of the cDNA coding for PPAR ⁇ 2 are obtained in the presence of the appropriate sense and antisense primers (Genset®, see Table 1).
  • the sizes of the PCR products are 307 base pairs (3T3-F442A cells) and 582 base pairs (human pre-adipocytes).
  • the appropriate controls reverse transcription and PCR amplification steps were performed.
  • PCR mixtures were subjected to 22 cycles (3T3-F442A cells) or 35 cycles (human preadipocytes) of denaturation amplification (2 minutes at 94 ° C), hybridization (1 minute at 60 ° C) and elongation (6 minutes). minutes at 72 ° C).
  • the PCR products are analyzed by agarose gel electrophoresis and visualized with ethidium bromide.
  • the level of expression of the adipogenic transcription factor PPAR ⁇ 2 is greatly decreased when the 3T3-F442A cells have been in contact with a solution of HE (FIG. 3A, result 2), of a solution of ACH (FIG. 3A, result 3 ), compared to control cells as evidenced by the intensity of the band (FIG. 3A, result 1).
  • the same results are obtained with the ACAE solution and the AOA solution.
  • the solutions of HE, ACH, ACAE or AOA according to the invention significantly inhibit the expression of the PPAR ⁇ 2 mRNA of human preadipocytes (FIG. 3B, the result 5 illustrating a treatment with a solution of HE, and the result 6 illustrating a treatment with ACH solution), compared to control human preadipocytes ( Figure 3B, result 4).
  • the essential oil of Allium sativum (HE) bulbs act at the level of the program of differentiation by decreasing the expression of the PPAR ⁇ 2 messenger.
  • HE, ACH, ACAE and AOA The ability of HE, ACH, ACAE and AOA to inhibit the differentiation process of human preadipocytes was determined by quantitative measurement of secreted leptin levels in the culture medium of the various assays. Indeed, leptin encoded by the ob gene , synthesized and secreted by adipocytes, is considered to be a late marker of the adipocyte differentiation process. In vitro, basal secretion of leptin progressively increases throughout the process of converting preadipocytes into mature adipocytes (MacDougald, OA et al, Proc Natl Acad Sci, USA, 92: 9034-9037, 1995).
  • leptin The effect on secretion of leptin from solutions of HE, ACH, ACAE and AOA was studied over a 9-day treatment period (human preadipocytes). Levels of leptin secreted by control and treated cells were measured in the various culture media, taken at 3, 6, and 9 days (days of renewal of the medium). Quantitative determination of released leptin levels is performed by ELISA assay (R & D Systems Europe) according to the supplier's instructions. The results are expressed as an average ⁇ standard deviation to the mean. The statistical comparisons are made by the Student's T test (p ⁇ 0.05 representing the threshold of significance).
  • Human preadipocytes cultured in the presence of HE, ACH, ACAE and AOA secrete in the culture medium amounts of leptin significantly lower than those detected in the culture medium of untreated control human preadipocytes.
  • the results obtained demonstrate that the solutions of HE, ACH, ACAE or AOA act at the level of the differentiation program and are capable of inhibiting the process of adipocyte conversion.

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EP01982561A 2000-10-31 2001-10-26 Composition anti-adipeuse a base d'extraits de bulbes d'ail et utilisations cosmetiques et therapeutiques Expired - Lifetime EP1335701B1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
FR0013973A FR2815865B1 (fr) 2000-10-31 2000-10-31 Utilisation d'une composition therapeutique ou cosmetique pour le traitement topique de la cellulite, et composition en comportant l'application
FR0013973 2000-10-31
PCT/FR2001/003325 WO2002036093A2 (fr) 2000-10-31 2001-10-26 Composition anti-adipeuse a base d'extraits de bulbes d'ail

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EP1335701A2 EP1335701A2 (fr) 2003-08-20
EP1335701B1 true EP1335701B1 (fr) 2006-06-14

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US (1) US6852343B2 (zh)
EP (1) EP1335701B1 (zh)
JP (1) JP4124648B2 (zh)
CN (1) CN1226996C (zh)
AT (1) ATE329575T1 (zh)
AU (2) AU1411102A (zh)
BR (1) BR0115009A (zh)
CA (1) CA2426284A1 (zh)
CY (1) CY1105267T1 (zh)
DE (1) DE60120744T2 (zh)
DK (1) DK1335701T3 (zh)
ES (1) ES2266280T3 (zh)
FR (1) FR2815865B1 (zh)
PT (1) PT1335701E (zh)
WO (1) WO2002036093A2 (zh)
ZA (1) ZA200303693B (zh)

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US7736661B1 (en) * 2000-03-07 2010-06-15 Avon Products, Inc Method of treating skin conditions
FR2831447B1 (fr) * 2001-10-26 2004-03-12 Innovaderm Absolues de bulbes d'allium sativum et utilisations a titre therapeutique ou cosmetique
US8273024B2 (en) * 2004-03-16 2012-09-25 Rita Kathleen Chew Ultrasound transmission gel
WO2006039285A2 (en) * 2004-09-30 2006-04-13 The University Of Georgia Research Foundation, Inc. Compositions and methods for inducing adipose tissue cell death
JP5049794B2 (ja) * 2005-06-10 2012-10-17 株式会社プライマリーセル 内臓前駆脂肪細胞から内臓脂肪細胞への分化を誘導する方法
WO2007089685A2 (en) * 2006-01-30 2007-08-09 University Of Georgia Research Foundation Compositions and methods for inducing adipose tissue cell death
JP2007230981A (ja) * 2006-03-03 2007-09-13 Hirosaki Univ ニンニク抽出物又はその分画・画分を有効成分とする心筋リモデリング抑制剤
CN108639792B (zh) * 2018-05-20 2023-11-24 成都鸿源锦程机器人有限公司 一种自动装车机器人的拨臂分料式装车机头

Family Cites Families (11)

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GB842404A (en) * 1958-06-06 1960-07-27 American Home Prod Skin treatment composition
GB1106551A (en) * 1964-03-18 1968-03-20 Roland Nessim Benhaim Pharmaceutically-active extracts of the nympheaceae
FR2628339B1 (fr) * 1988-03-14 1991-04-26 Rhone Poulenc Chimie Emulsions aqueuses stables d'huiles essentielles
FR2660312A1 (fr) * 1990-03-30 1991-10-04 Ardeval Laboratoires Compositions a base de plantes fraiches ou seches et leurs applications.
TW205068B (zh) * 1991-03-08 1993-05-01 Takeda chemical industry co ltd
JPH04338336A (ja) * 1991-05-15 1992-11-25 Riken Health Kk 抗肥満剤
FR2693371B1 (fr) * 1992-07-08 1994-09-02 Gisele Melin Composition pour crème de beauté antirides.
EP0923937B1 (en) * 1997-12-08 2004-03-10 Council of Scientific and Industrial Research A herbal formulation useful as a therapeutic and cosmetic application for the treatment of general skin disorders
KR100525841B1 (ko) * 1997-12-27 2005-12-21 주식회사 엘지생활건강 양파 추출물을 함유하는 슬리밍 화장품 조성물
DE19858670C2 (de) * 1998-12-18 2001-07-12 Heinz Kasik Mittel zur Pflege von Haut, Nägeln und Haaren und zur Förderung des Haarwachstums
FR2804319B1 (fr) 2000-01-28 2002-10-25 Clarins Lab Composition cosmetique amincissante comprenant en tant qu'agent actif un extrait de plante contenant de l'anp

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BR0115009A (pt) 2003-09-30
ATE329575T1 (de) 2006-07-15
AU2002214111B9 (en) 2006-11-09
DK1335701T3 (da) 2006-10-16
DE60120744D1 (de) 2006-07-27
ES2266280T3 (es) 2007-03-01
US6852343B2 (en) 2005-02-08
CA2426284A1 (fr) 2002-05-10
AU1411102A (en) 2002-05-15
FR2815865B1 (fr) 2003-03-21
ZA200303693B (en) 2004-08-13
DE60120744T2 (de) 2007-06-14
WO2002036093A3 (fr) 2003-01-09
EP1335701A2 (fr) 2003-08-20
JP2004522704A (ja) 2004-07-29
JP4124648B2 (ja) 2008-07-23
WO2002036093A2 (fr) 2002-05-10
CN1226996C (zh) 2005-11-16
CN1487822A (zh) 2004-04-07
US20020146474A1 (en) 2002-10-10
CY1105267T1 (el) 2010-03-03
AU2002214111B2 (en) 2006-06-08
FR2815865A1 (fr) 2002-05-03
PT1335701E (pt) 2006-10-31

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